Volume 26, Issue 1 (1-2022)                   ibj 2022, 26(1): 85-90 | Back to browse issues page

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Hezarjaribi N, Fazeli M R. A Simplified Process for Purification and Refolding of Recombinant Human Interferon-α2b. ibj. 2022; 26 (1) :85-90
URL: http://ibj.pasteur.ac.ir/article-1-3476-en.html
Background: Interferon α-2b is a vital biotherapeutic produced through the recombinant DNA technology in E. coli. The recombinant IFN-α2b normally appears as intercellular IBs, which requires intensive refolding and purification steps. Method: Purification of IFN-α2b from solubilized IB was performed using two-phase extraction. To optimize refolding conditions, the effects of pH and different additives, including cysteine, cystine, urea, glycerol, Triton X-100, NaCl, and arginine, were investigated. Optimal refolding buffer (0.64 mM of urea, 5.57 mM of cysteine ​​, and 1.8 mM of cystine) was obtained using RSM. The refolding process was performed by an optimized refolding buffer in the dilution and fed-batch refolding method at different protein concentrations (25-1000 µg/mL). Result:  At a final protein concentration of 500 µg/mL, the fed-batch refolding method yielded in a biological activity of 2.24 × 108 IU/mg, which was nearly twice that of dilution method. Conclusion: Fed-batch refolding method resulted in the biologically active IFN-α2b with high purity, which can be used for research and industrial purposes.
Type of Study: Full Length | Subject: Pharmaceutical Biotechnology

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